Chemical and immunological characterization of the extracellular galactomannan of Aspergillus fumigatus

Latgé, Jean‐Paul; Kobayashi, Hidemitsu; Debeaupuis, J P; Diaquin, Michel; Sarfati, J.; Wieruszeski, J.-M.; Parra, Enrique; Bouchara, Jean Philippe; Fournet, Bernard

doi:10.1128/iai.62.12.5424-5433.1994

Cited by 286 publications

(147 citation statements)

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“…The EBA2 was raised against extracellular galactomannan (GM) from the pathogenic fungus Aspergillus fumigatus [19]. GM was subsequently characterised and shown to be composed of a linear mannan core with side chains of up to ¢ve L-1,5-linked Galf units [3]. The mannan core is not antigenic.…”

Section: Resultsmentioning

confidence: 99%

β-Galactofuranoside glycoconjugates on conidia and conidiophores of Aspergillus niger

Wallis¹

2001

FEMS Microbiology Letters

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Galactose in the furanoic conformation appears to be limited to bacteria and lower eukaryotes. Galactofuranoic (Galf)-containing glycoconjugates that occur in organisms pathogenic or allergenic to man are frequently antigenic and immunodominant. We have used an immunochemical approach, employing a monoclonal antibody that recognises Galf epitopes, to investigate the presence of Galf-containing glycoconjugates within conidia and conidiophores of Aspergillus niger. ELISA and immunofluorescence microscopy indicated that specific and saturable binding sites were found on both. Inhibition studies confirmed that this binding was to Galf-containing glycoconjugates. Interestingly, the conidiophore heads were particularly rich in these glycoconjugates. Western blotting identified a Galf glycoprotein of 1502 00 kDa from disrupted conidia. ß

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Section: Resultsmentioning

confidence: 99%

β-Galactofuranoside glycoconjugates on conidia and conidiophores of Aspergillus niger

Wallis¹

2001

FEMS Microbiology Letters

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“…Enzyme purification. Mycelial culture of A. fimigatus, strain CBS 143-89 and preparation of cell wall autolysate were carried out as previously described (LatgC et al, 1994;Hartland et al, 1996).…”

Section: Methodsmentioning

confidence: 99%

Purification and Characterization of an Endo‐1,3‐β‐Glucanase from Aspergillus fumigatus

Fontaine

Hartland

Beauvais

et al. 1997

European Journal of Biochemistry

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An endo-I ,3-p-glucanase was purified from a cell wall autolysate of Aspcvgillus ,fumigatus. This pglucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detector for product analysis, it was shown that the endoglucanase hydrolysed exclusively linear 1,3-P-glucan chains, had an optimum pH of 7.0 and an optimum temperature of 60°C. A substrate kinetic study gave a K,, value of 0.3 mg/ml for soluble (laminarin and laminari-oligosaccharides) and 1 .18 mg/ml for insoluble (curdlan) 1,3-P-glucan. Laminar-oligosaccharide degradation, analysed by HPLC, showed that the endoglucanase bind to the subtrate at several positions and suggested that the active site of the enzyme recognized five glucose units linked by a 1,3-1 bond. The association of the present endo-1,3-P-glucanase with the cell wall of A. fumigatus suggests a putative role for this enzyme during cell-wall morphogenesis.Keywords: endo-I ,3-P-glucanase ; cell wall : Aspergillus furnigutus; filamentous fungus.1 ,3-P-Glucanases are classified as exo-l,3-/$glucanase (1,3-P-glucan glucohydrolase) and endo-1,3-P-glucanase (1,3-1-glucan glucanohydrolase). Endoglucanase activities cleave inside a glucan chain in a more or less random fashion: while the exoglucanase activities release glucose residues from the non-reducing end. Many yeast and filamentous fungi produce both exocellular and cell-wall-associated exo-p-glucanases and endo-8-glucanases (Notario, 1982;Hien and Fleet, 1983 ;Copa-Patino, et al. 1989;Stahmann et al., 1993). 1,3-p-Glucan is a predominant polysaccharide of the fungal cell wall, and is thought to be responsible for the shape and rigidity of the wall (Cabib et al., 1982;Fleet, 1991). It has been suggested that 1,3-p-glucanase activities are essential to the continuous rearrangement of the wall 1,3-P-glucans during fungal growth (Cabib et al., 1988;Wessels, 1988). In the filamentous fungus Coprinus cinereus, endo-p-glucanases seem to play an essential role in stipe elongation (Kamada et al., 1985). In Sacchuronzyces cerevisiae, the production of exo-P-glucanases is growth-associated and cellcycle regulated (del Rey et al., 1979;Larriba et al., 1984;Jiang et al., 1995), suggesting that their activities are required at very specific stages during morphogenesis. For example, bud initiation, cell expansion, cell conjugation and sporulation are characterised by a higher activity of particular 1,3-~-glucanases (del Rey et al., 1982; Cenamor et al., 1987). However, gene disruption of yeast exo-p-glucanase activities does not modify the cellular growth (Larriba et al., 1995).Correspondence to T. Fontaine, lnstitut Pasteur, Laboratoire des Aspergillus, 25 rue du Docteur E. Roux, F-75724 Paris cedex 15, FranceAbbreviafions. Abz(OH)ONHNHZ, pamino-hydroxybenzoic acid hydrazide; Con-A, concanavalin A lcctin; dp, degree of polymerisation; Gn, oligosaccharide containing n 1,3-P-linked glucose residues ; G,,r, reduced oligosacchari...

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“…b-Galactofuranosyl residues have been shown to be a component of the galactomannan produced by Aspergillus spp. [31,41], the O-linked glycan of A. niger GAM [20,23], invertase secreted by A. nidulans [27] and part of other fungal glycoproteins [32,42,43]. Takayanagi et al [9] also observed the, to date, novel structure of a-linked Galf residues on a different a-galactosidase produced by A. niger.…”

Section: Biological Significancementioning

confidence: 99%

Galactofuranoic‐oligomannose N‐linked glycans of α‐galactosidase A fromAspergillus niger

Wallis

Easton

Jolly³

et al. 2001

European Journal of Biochemistry

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Extracellular a-galactosidase A was purified from the culture filtrate of an over-producing strain of Aspergillus niger containing multiple copies of the encoding aglA gene under the control of the glucoamylase (glaA) promoter. Endoglycosidase digestion followed by SDS/PAGE, lectin and immunoblotting suggested that glycosylation accounted for < 25% of the molecular size of the purified protein. Monosaccharide analysis showed that this was composed of N-acetyl glucosamine, mannose and galactose. Mild acid hydrolysis, mild methanolysis, immunoblotting and exoglycosidase digestion indicated that the majority of the galactosyl component was in the furanoic conformation (b-d-galactofuranose, Galf). At least 20 different N-linked oligosaccharides were fractionated by high-pH anion-exchange chromatography following release from the polypeptide by peptide-N-glycosidase F. The structures of these were subsequently determined by fast atom bombardment mass spectrometry to be a linear series of Hex 7226 HexHAc 2 . Indicating that oligosaccharides from GlcNAc 2 Man 7 , increasing in molecular size up to GlcNAc 2-Man 24 were present. Each of these were additionally substituted with up to three b-Galf residues. Linkage analysis confirmed the presence of mild acid labile terminal hexofuranose residues. These results show that filamentous fungi are capable of producing a heterogeneous mixture of high molecular-size N-linked glycans substituted with galactofuranoic residues, on a secreted glycoprotein.

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Chemical and immunological characterization of the extracellular galactomannan of Aspergillus fumigatus

Cited by 286 publications

References 45 publications

β-Galactofuranoside glycoconjugates on conidia and conidiophores of Aspergillus niger

β-Galactofuranoside glycoconjugates on conidia and conidiophores of Aspergillus niger

Purification and Characterization of an Endo‐1,3‐β‐Glucanase from Aspergillus fumigatus

Galactofuranoic‐oligomannose N‐linked glycans of α‐galactosidase A fromAspergillus niger

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